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Background/aim: Our study investigated the susceptibility rate of ceftazidime-avibactam and the risk factors associated with its resistance by analyzing gram-negative bacteria isolated from various patient samples. Materials and methods: Between March and November 2020, 1119 gram-negative bacteria strains were isolated from patient samples in Acıbadem Healthcare Group hospitals; ceftazidime-avibactam susceptibility results were evaluated using a 10/4µg (Oxoid, UK) disc and evaluated according to Eucast 2020 recommendations. Patient and isolate characteristics that could be risk factors were retrospectively investigated and statistically analyzed using SPSS 25.0. Results: Male patients made up 52% (n = 581) of the study's total patient population, and they averaged 55.5 ± 24.9 years old. Of 1119 gram-negative strains culture and antibiogram, 1023 (91.4%) were sensitive to ceftazidime-avibactam. An increased risk of resistance was observed with female gender (OR = 2.29; CI 95% [1.45-3.61]; p < 0.05), Pseudomonas aeruginosa (OR = 1.67, CI 95% [1.03-2.7]; p < 0.05), the presence of multidrug-resistance (MDR) (OR = 4.07, CI 95% [2.47-6.7]; p < 0.05) pandrug-resistance (PDR) (OR = 12, (CI) 95% [9.9-14.7]]; p < 0.05) and admission to intensive care unit (ICU) (OR = 1.89, CI 95% [1.22-2.93]; p < 0.05). Conclusion: The resistance rate of ceftazidime-avibactam was found to be 8.6%, and it was thought that resistant strains produced metallo-ß-lactamase (MBL) type carbapenemase. Risk factors were female gender, Pseudomonas aeruginosa, MDR, PDR, and admission to ICU. Therefore, studying the ceftazidime-avibactam susceptibility test together with gram-negative bacteria identification, especially in groups at risk for resistance, is one of the important factors that can positively affect the success of treatment. [ABSTRACT FROM AUTHOR]

In this paper, the kinetics of p53 in two cell lines with different degrees of sensitivity to chemotherapeutic drugs is studied. There is much research that has explored the p53 oscillation, but there are few comparisons between cells that are sensitive to drug treatment and those that are not. Here, the kinetics of the p53 system between etoposide-sensitive and etoposide-resistant cell lines in response to different drug doses and different protein synthesis time delays are studied and compared. First, the results showed that time delay is an important condition for p53 oscillation by producing Hopf bifurcation in both the etoposide-sensitive and etoposide-resistant cells. If the protein synthesis time delays are zero, the system cannot oscillate even the dose of the drug increases. Second, the time delay required for producing sustained oscillation in sensitive cells is shorter than the drug-resistant cells. In addition, the p53-Wip1 negative feedback loop in drug-resistant cells is relatively highly strengthened than the drug-sensitive cells. To sum up, p53 oscillation is controlled by time delay, drug dose, and the coupled negative feedback network including p53-mdm2 and p53-wip1. Moreover, in the two different types of cells, the control mechanisms are similar, but there are also differences. [ABSTRACT FROM AUTHOR]

Breast cancer is a heterogeneous disease, and the survival rate of patients with breast cancer strongly depends on their stage and clinicopathological features. Chemoradiation therapy is commonly employed to improve the survivability of patients with advanced breast cancer. However, the treatment process is often accompanied by the development of drug resistance, which eventually leads to treatment failure. Metabolism reprogramming has been recognized as a mechanism of breast cancer resistance. In this study, we established a doxorubicin-resistant MCF-7 (MCF-7-D500) cell line through a series of long-term doxorubicin in vitro treatments. Our data revealed that MCF-7-D500 cells exhibited increased multiple-drug resistance, cancer stemness, and invasiveness compared with parental cells. We analyzed the metabolic profiles of MCF-7 and MCF-7-D500 cells through liquid chromatography–mass spectrometry. We observed significant changes in 25 metabolites, of which, 21 exhibited increased levels (>1.5-fold change and p < 0.05) and 4 exhibited decreased levels (<0.75-fold change and p < 0.05) in MCF-7 cells with doxorubicin resistance. These results suggest the involvement of metabolism reprogramming in the development of drug resistance in breast cancer, especially the activation of glycolysis, the tricarboxylic acid (TCA) cycle, and the hexamine biosynthesis pathway (HBP). Furthermore, most of the enzymes involved in glycolysis, the HBP, and the TCA cycle were upregulated in MCF-7-D500 cells and contributed to the poor prognosis of patients with breast cancer. Our findings provide new insights into the regulation of drug resistance in breast cancer, and these drug resistance-related metabolic pathways can serve as targets for the treatment of chemoresistance in breast cancer. [ABSTRACT FROM AUTHOR]

Identification and phenotypic drug-susceptibility testing for mycobacteria are time-consuming and challenging but essential for managing mycobacterial infections. Next-generation sequencing (NGS) technologies can increase diagnostic speed and quality, but standardization is still lacking for many aspects (e.g., unbiased extraction, host depletion, bioinformatic analysis). Targeted PCR approaches directly on sample material are limited by the number of targets that can be included. Unbiased shotgun metagenomics on direct material is hampered by the massive amount of host DNA, which should be removed to improve the microbial detection sensitivity. For this reason, we developed a method for NGS-based diagnosis of mycobacteria directly from patient material. As a model, we used the non-tuberculous mycobacterium (NTM) Mycobacterium abscessus. We first compared the efficiency of three different DNA extraction kits for isolating DNA (quality and concentration). The two most efficient kits were then used in a follow-up study using artificial sputum. Finally, one extraction kit was selected and further evaluated for DNA isolation from a patients’ sputum mixture spiked with M. abscessus at three concentrations (final concentrations 108, 107, 106  CFU/ml). The spiked sputum samples were processed with and without saponin treatment (ST) in combination with DNAse treatment prior to bacterial DNA extraction to evaluate the recovery of bacteria and depletion of host DNA by PCR and Illumina sequencing. While Ct values of the qPCR targeting mycobacterial ITS DNA remained rather stable, Ct values in the qPCR targeting the human β-actin gene increased by five Ct values in ST samples. In subsequent Illumina sequencing, a decrease of 89% of reads mapped to the human genome was observed in ST samples. The percentage of reads mapped to M. abscessus (108 CFU/ml) increased by 89%, and the sequencing depth increased two times when undergoing ST. In conclusion, the sensitivity of M. abscessus detection in artificial sputum was increased using a saponin pre-treatment step. The saponin followed by the DNase I treatment approach could be efficiently applied to detect and characterize mycobacterial infections, including tuberculosis, directly from sputum. [ABSTRACT FROM AUTHOR]

Background Anthrax is endemic to many countries, including the United States. The causative agent, Bacillus anthracis, poses a global bioterrorism threat. Without effective antimicrobial postexposure prophylaxis (PEPAbx) and treatment, the mortality of systemic anthrax is high. To inform clinical guidelines for PEPAbx and treatment of B. anthracis infections in humans, we systematically evaluated animal anthrax treatment model studies. Methods We searched for survival outcome data in 9 scientific search engines for articles describing antimicrobial PEPAbx or treatment of anthrax in animals in any language through February 2019. We performed meta-analyses of efficacy of antimicrobial PEPAbx and treatment for each drug or drug combination using random-effects models. Pharmacokinetic/pharmacodynamic relationships were developed for 5 antimicrobials with available pharmacokinetic data. Monte Carlo simulations were used to predict unbound drug exposures in humans. Results We synthesized data from 34 peer-reviewed studies with 3262 animals. For PEPAbx and treatment of infection by susceptible B. anthracis , effective monotherapy can be accomplished with fluoroquinolones, tetracyclines, β-lactams (including penicillin, amoxicillin-clavulanate, and imipenem-cilastatin), and lipopeptides or glycopeptides. For naturally occurring strains, unbound drug exposures in humans were predicted to adequately cover the minimal inhibitory concentrations (MICs; those required to inhibit the growth of 50% or 90% of organisms [MIC50 or MIC90]) for ciprofloxacin, levofloxacin, and doxycycline for both the PEPAbx and treatment targets. Dalbavancin covered its MIC50 for PEPAbx. Conclusions These animal studies show many reviewed antimicrobials are good choices for PEPAbx or treatment of susceptible B. anthracis strains, and some are also promising options for combating resistant strains. Monte Carlo simulations suggest that oral ciprofloxacin, levofloxacin, and doxycycline are particularly robust choices for PEPAbx or treatment. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) is a Gram-positive pathogenic bacterium, which persistently colonizes the anterior nares of approximately 20--30% of the healthy adult population, and up to 60% is intermittently colonized. With the misuse and overuse of antibiotics, large-scale drug-resistant bacteria, including methicillin-resistant S. aureus (MRSA), have been appeared. MRSA is among the most prevalent pathogens causing community-associated infections. Once out of control, the number of deaths caused by antimicrobial resistance may exceed 10 million annually by 2050. Antimicrobial peptides (AMPs) are regarded as the best solution, for they are not easy to develop drug resistance. Based on our previous research, here we designed a new antimicrobial peptide named GW18, which showed excellent antimicrobial activity against S. aureus, even MRSA, with the hemolysis less than 5%, no cytotoxicity, and no acute toxicity. Notably, administration of GW18 significantly decreased S. aureus infection in mouse model. These findings identify GW18 as the ideal candidate against S. aureus infection. [ABSTRACT FROM AUTHOR]

Background: Aspirin (ASA) has been reported to have an antitumor effect but the role of ASA in the prevention and treatment of breast cancer (BC) is still controversial. This study aimed to identify clinical effectiveness of ASA in the treatment of BC and explore the antitumor target proteins of ASA that may be involved in overcoming tamoxifen resistance in estrogen receptor (ER)-positive BC cells. Materials and Methods: Randomized controlled trials (RCTs) of ASA in the treatment of BC were queried from the databases, including PubMed, Web of Science, Cochrane Library, WanFang, and Chinese National Knowledge Infrastructure. According to the quality standard recommended in the Newcastle-Ottawa Scale (NOS), the outcome indexes were analyzed by RevMan 5.3 and Stata 12.0 software. Cell culture experiments were performed to explore the effect of tamoxifen combined with ASA on the proliferation of ER-positive BC cell lines MCF-7 and MCF-7/TAM. Cell cytotoxicity was determined by the 3-(4, 5-di-2-yl)-2, 5-ditetrazolium bromide (MTT) assay. A quantitative proteomic analysis was conducted between the control and experimental groups to identify differentially expressed proteins (DEPs). Subsequently, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used for bioinformatic analysis of DEPs. The protein expression in patients of ER-positive BC was analyzed by immunochemistry (IHC). Results: Nine RCTs including 162,381 patients were selected for this study. The meta-analysis revealed that daily ASA use, as compared with its non-use, was associated with a decreased risk of BC death: relative risk (RR) = 0.83%, 95% CI [0.73, 0.94], Z = 2.89, P = 0.004 (P < 0.05). Cell culture experiments showed that tamoxifen combined with ASA can drastically inhibit the cell growth of MCF-7 and MCF-7/TAM cells more than the administration of tamoxifen alone (P < 0.05). Fifty-seven DEPs were up-regulated, while eighty-five DEPs were down-regulated in MCF-7/TAM cells after the ASA combination treatment. Several GO terms were significantly enriched, such as neutrophil degranulation, retinal metabolic process, sterol biosynthetic process, and prostaglandin metabolic process. KEGG pathway enrichment analysis also verified three associated pathways including metabolic pathways, chemical carcinogenesis-reactive oxygen species, and biosynthesis of amino acids. In ER-positive BC patients with Ki67 > 20%, the positive expression of one significantly DEP MYC was much higher than in BC patients with Ki67 = 20% (40.91% vs. 12.50%, P < 0.05). There was no significant difference in MYC protein expression among the other subgroups (P > 0.05). Conclusions: Our results show that ASA has a clinical value in the treatment of BC. ASA could overcome tamoxifen resistance in ER-positive BC cells through some key proteins, which may be potential therapeutic targets for patients with tamoxifen resistance. [ABSTRACT FROM AUTHOR]

Tuberculosis, an infectious disease, is one of the leading causes of death worldwide. Drug-resistant tuberculosis exacerbates its threat. Despite long-term and costly treatment with second-line drugs, treatment failure rates and mortality remain high. Therefore, new strategies for developing new drugs and improving the efficiency of existing drug treatments are urgently needed. Our research team reported that PPs, a new class of potential anti-tuberculosis drug candidates, can inhibit the growth of drug-resistant Mycobacterium tuberculosis. Here, we report a synergistic effect of PPs with ethionamide (ETH), one of the second-line drugs, as a result of further research on PPs. While investigating gene expression changes based on microarray and 2DE (two-dimensional gel electrophoresis), it was found that PPs induced the greatest overexpression of Rv0560c in M. tuberculosis. Based on this result, a protein microarray using Rv0560c protein was performed, and it was confirmed that Rv0560c had the highest interaction with EthR, a repressor for EthA involved in activating ETH. Accordingly, a synergistic experiment was conducted under the hypothesis of increased susceptibility of ETH to M. tuberculosis by PPs. As a result, in the presence of 0.5× MIC PPs, ETH showed a growth inhibitory effect on drug-sensitive and -resistant M. tuberculosis even at a much lower concentration of about 10-fold than the original MIC of ETH. It is also suggested that the effect was due to the interaction between PPs and Rv2887, the repressor of Rv0560c. This effect was also confirmed in a mouse model of pulmonary tuberculosis, confirming the potential of PPs as a booster to enhance the susceptibility of M. tuberculosis to ETH in treating drug-resistant tuberculosis. However, more in-depth mechanistic studies and extensive animal and clinical trials are needed in the future. [ABSTRACT FROM AUTHOR]

Non-fermenting gram-negative bacteria (NFGNB) frequently exhibit drug resistance. The purpose of this study was to determine the drug resistance pattern among the NFGNB isolates causing respiratory tract infections (RTI s). A retrospective analysis of the antimicrobial susceptibility pattern of non-fermenters causing RTI s over four years (2016-2019) was done and the change in drug resistance pattern was studied. A total of 653 cases were obtained that included 191 (29.2%) Moraxella catarrhalis, 283 (43.3%) Pseudomonas aeruginosa, and 132 (20.2%) Acinetobacter baumannii, 47 (7.2%) Stenotrophomonas maltophilia isolates. A higher resistance (82.6%) was observed for piperacillin-tazobactam and cefpirome, followed by imipenem (79.5%) and ciprofloxacin (76.5 %) for A. baumannii isolates. A sharp decline in resistance pattern for piperacillin, cefpirome, Imipenem and cefoperazone-sulbactam in 2019 and an increasing resistance to gentamycin and ciprofloxacin were noted. Among P. aeruginosa isolates, 94% aztreonam and 83.4% cefoperazone-sulbactam resistance were detected. There was an increased resistance for cefpirome and piperacillin and a decreased resistance for Imipenem was recorded in 2019. In cases of M. catarrhalis, 22.51% of isolates were resistant to ciprofloxacin, followed by erythromycin (18.32%) and tetracycline (17.80 %). S. maltophilia showed a 100% sensitivity for cotrimoxazole and 2.1% resistance for ciprofloxacin. A constantly changing antibiotic-resistant pattern of non-fermenters compels for a continuous update of drug-resistant trends through a longitudinal surveillance program in different geographical areas. [ABSTRACT FROM AUTHOR]

Fungal disease affects over a billion people worldwide. Naamidine A inhibits the growth of diverse fungal pathogens through an unknown mechanism. Here, we show that the supplementation of medium with excess zinc abolishes the antifungal activity of naamidine A. Furthermore, we highlight that naamidine A has in vitro activity against terbinafine-resistant Trichophyton spp . and in vivo efficacy in a mouse model of dermatomycosis caused by T. mentagrophytes, highlighting its therapeutic potential as a topical treatment., Competing Interests: L.E.C. and L.W. are co-founders and shareholders in Bright Angel Therapeutics, a platform company for the development of novel antifungal therapeutics. L.E.C. is a Science Advisor for Kapoose Creek, a company that harnesses the therapeutic potential of fungi.

Tuberous sclerosis Complex (TSC) is a genetic disorder characterized by multisystem involvement, with epilepsy affecting 80-90% of patients, often beginning in infancy. Early-life seizures in TSC are associated with poor neurodevelopmental outcomes, underscoring the importance of timely and effective management. This review explores the evolving treatment landscape for TSC-associated seizures in young children, focusing on three recently approved or license-expanded therapies: vigabatrin, everolimus, and cannabidiol. The efficacy and safety profiles of these treatments are examined based on clinical trials and real-world evidence, with a focus on their use in treating seizures in young children. The preemptive use of vigabatrin in clinical studies has also been carefully reviewed. A treatment algorithm is proposed, emphasizing early diagnosis, prompt initiation of appropriate therapy, and a stepwise approach to managing both infantile spasms and focal seizures. The algorithm incorporates these newer therapies alongside traditional antiseizure medications and non-pharmacological approaches. Challenges in optimizing treatment strategies, minimizing side effects, and improving long-term outcomes are discussed. This review aims to guide clinicians in navigating the complex landscape of early-life seizures associated with TSC, ultimately striving for improved seizure control and better developmental outcomes in this vulnerable population., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Debopam Samanta collected the data, analyzed and interpreted them, and drafted and revised the manuscript for intellectual content.He declares no potential conflicts of interest with respect to this article's research, authorship, and/or publication.]., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Purpose: To provide consensus-based recommendations for use of vagus nerve stimulation (VNS) therapy in the management of pediatric epilepsy., Methods: Delphi methodology with two rounds of online survey was used to build consensus. A steering committee developed 43 statements related to pediatric epilepsy and the use of VNS therapy, which were evaluated by a panel of 12 neurologists/neurosurgeons with expertise in pediatric epilepsy, who graded their agreement with each statement on a scale of 1 ("I do not agree at all") to 5 ("I strongly agree"). For each statement, consensus was established if ≥70% of the agreement scores were 4 or 5 and <30% were 1 or 2 in the final survey., Results: Twenty-four statements regarding the need for seizure reduction in pediatric epilepsy, the recommended treatment algorithm, the benefits and safety of VNS therapy, management of side effects of VNS therapy, patient selection for VNS therapy, and the use, dosing, and titration of VNS therapy achieved consensus. VNS and other neuromodulation therapies should be considered for pediatric patients with drug-resistant epilepsy who are not candidates for resective surgery, or who do not remain seizure free after resective surgery. When VNS therapy is initiated, the target dose range should be achieved via the fastest and safest titration schedule for each patient. Scheduled programming can be helpful in dose titration., Conclusion: The expert consensus statements represent the panelists' collective opinion on the best practice use of VNS therapy to optimize outcomes in the management of pediatric epilepsy., Competing Interests: Declaration of competing interest James W. Wheless is a consultant for BioMarin, Epiwatch, Neurelis, Neuro Event Labs, Jazz, and Takeda; has received research funding from LivaNova, Jazz, Neurelis, Neuro Event Labs, Shainberg Foundation, SKLSI, TSC Alliance, and UCB; and serves on the speaker's bureau for BioMarin, Jazz, Neurelis, SKLSI, and UCB. Jeffrey S. Raskin is a clinical advisory board member for Synergia, a consultant for Blackrock Neurotech, Iota, and Brainlab, and President of SMERF, Inc 501(c)(3). Anthony L. Fine has no financial interests to disclose. Kelly Knupp has consulted for UCB, Stoke, Encoded, Epygenix, Longboard and Biocodex; has received research funding from Eisai, Stoke, and UCB; and serves on a Data and Safety Monitoring Board for Jazz and Epygenix. John Schreiber is a clinical advisory board member for Stoke Therapeutics and Longboard Pharmacueticals; a consultant for Denali Therapeutics and Neurocrine; receives research funding to his institution from UCB, Neurocrine, and Stoke Therapeutics; and serves on the speaker bureau for UCB and Marinus. Adam P. Ostendorf receives research funding from NINDS and the Nationwide Children's Hospital Foundation. Gregory W. Albert has no financial interests to disclose. Eric H. Kossoff has received consultant fees from LivaNova (not for this study), Simply Good Foods, Nutricia, Vitaflo, Cerecin, and Bloom Science. He is on the data safety monitoring board for a study from NeuroPace. Joseph R. Madsen is a consultant and scientific advisory board member for BioNaut Laboratories and Anuncia Medical. Prakash Kotagal is a consultant for Biocodex, Inc., LivaNova PLC, SK Life Sciences, and Takeda Pharmaceuticals. Adam L. Numis has no financial interests to disclose. Nisha Gadgil has no financial interests to disclose. Deborah L. Holder is a speaker for LivaNova. Elizabeth A. Thiele is a consultant for Jazz, UCB, Biocodex, LivaNova, Pyros, SKLS, Stoke Therapeutics, Takeda, Marinus, and Nobelpharma. She has received research funding from Jazz, UCB, and Biocodex. George M. Ibrahim has received research funding and consulting fees from LivaNova (not for this study). He also receives consulting and advisory fees from Medtronic Inc and Synergia Inc., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Antimicrobial peptides (AMP) have emerged as promising candidates for addressing the clinical challenges posed by the rapid evolution of antibiotic-resistant microorganisms. Brevinins, a representative frog-derived AMP family, exhibited broad-spectrum antimicrobial activities, attacking great attentions in previous studies. However, their strong haemolytic activity and cytotoxicity, greatly limit their further development. In this work, we identified and characterised a novel brevinin-1 peptide, brevinin-1pl, from the skin secretions of the northern leopard frog, Rana pipiens . Like many brevinins, brevinin-1pl also displayed strong haemolytic activity, resulting in a lower therapeutic index. We employed several bioinformatics tools to analyse the structure and potential membrane interactions of brevinin-1pl, leading to a series of modifications. Among these analogues, des-Ala 16 -[Lys 4 ]brevinin-1pl exhibited great enhanced therapeutic efficacy in both in vitro and in vivo tests, particularly against some antibiotics-resistant Escherichia coli strains. Mechanistic studies suggest that des-Ala 16 -[Lys 4 ]brevinin-1pl may exert bactericidal effects through multiple mechanisms, including membrane disruption and DNA binding. Consequently, des-Ala 16 -[Lys 4 ]brevinin-1pl holds promise as a candidate for the treatment of drug-resistant Escherichia coli infections., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Approximately 30% of people with epilepsy will be refractory. This manuscript reviews current evidencebased non-surgical treatment modalities for pediatric refractory epilepsy, including pharmacological and dietary strategies.

Despite the regular discovery of new molecules, one-third of epileptic patients are resistant to antiepileptic drugs. Only a few can benefit from resective surgery, the current gold standard. Although effective in 50-70% of cases, this therapy remains risky, costly, and can be associated with long-term cognitive or neurological side effects. In addition, patients are increasingly reluctant to have a craniotomy, emphasizing the need for new less invasive therapies for focal drug-resistant epilepsies. Here, we review different minimally invasive approaches already in use in the clinic or under preclinical development to treat drug-resistant epilepsies. Localized thermolesion of the epileptogenic zone has been developed in the clinic using high-frequency thermo-coagulations or magnetic resonance imaging-guided laser or ultrasounds. Although less invasive, they have not yet significantly improved the outcomes when compared with resective surgery. Radiosurgery techniques have been used in the clinic for the last 20years and have proven efficiency. However, their efficacy is not better than resective surgery, and various side effects have been reported as well as the potential risk of sudden unexpected death associated with epilepsy. Recently, a new strategy of radiosurgery has emerged using synchrotron-generated X-ray microbeams: microbeam radiation therapy (MRT). The low divergence and high-flux of the synchrotron beams and the unique tolerance to MRT by healthy brain tissues, allows a precise targeting of specific brain regions with minimal invasiveness and limited behavioral or functional consequences in animals. Antiepileptic effects over several months have been recorded in animal models, and histological and synaptic tracing analysis suggest a reduction of neuronal connectivity as a mechanism of action. The possibility of transferring this approach to epileptic patients is discussed in this review., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Since 2022, Europe has had 4 cases of extensively drug-resistant Neisseria gonorrhoeae, sequence type 16406, that is resistant to ceftriaxone and highly resistant to azithromycin. We report 2 new cases from France in 2023 involving strains genetically related to the 4 cases from Europe as well as isolates from Cambodia.

This study sought to investigate the distribution, antimicrobial resistance rate, and bacterial co-infection frequency of non-tuberculous mycobacteria (NTM) in a single center in Incheon, South Korea. A total of 8,258 specimens submitted for tuberculosis (TB)/NTM real-time PCR tests during the years 2015 to 2020 were retrospectively reviewed. In total, 296 specimens (3.6%) were NTM positive, and the positivity increased from 2.5% (30/1,209) in 2015 to 3.8% (66/1,740) in 2020. Of 296 NTM specimens, 54.7% (162/296) were identified as the Mycobacterium avium complex (MAC) followed by the Mycobacterium abscessus complex (MABC) 20.9% (62/296), M. fortuitum 6.4% (19/296) and M. flavescens 3.4% (10/296). Of the NTM-positive specimens, 76.7% (227/296) were tested for drug resistance. The results showed multidrug-resistant NTM in 40.1% (91/227) and extensively drug-resistant NTM in 59.9% (136/227) of these specimens. Of the 145 isolates taken for bacterial culture, bacteria/fungi co-infection with NTM accounted for 43.4% (63/145), in which the most common bacterial species was Klebsiella pneumonia (23.8%, 15/63). This study is the first report on the distribution and antimicrobial resistance of NTM in Incheon. As the proportion of NTM infections increases, active treatment and thorough infection control are required for effective management.

Ovarian cancer is one of the most lethal gynecological malignancies. Recurrence or acquired chemoresistance is the leading cause of ovarian cancer therapy failure. Overexpression of ATP-binding cassette subfamily B member 1 (ABCB1), commonly known as P-glycoprotein, correlates closely with multidrug resistance (MDR). However, the mechanism underlying aberrant ABCB1 expression remains unknown. Using a quantitative high-throughput combinational screen, we identified that terfenadine restored doxorubicin sensitivity in an MDR ovarian cancer cell line. In addition, RNA-seq data revealed that the Ca2+-mediated signaling pathway in the MDR cells was abnormally regulated. Moreover, our research demonstrated that terfenadine directly bound to CAMKIID to prevent its autophosphorylation and inhibit the activation of the cAMP-responsive element-binding protein 1 (CREB1)-mediated pathway. Direct inhibition of CAMKII or CREB1 had the same phenotypic effects as terfenadine in the combined treatment, including lower expression of ABCB1 and baculoviral IAP repeat-containing 5 (BIRC5, also known as survivin) and increased doxorubicin-induced apoptosis. In this study, we demonstrate that aberrant regulation of the Ca2+-mediated CAMKIID/CREB1 pathway contributes to ABCB1 over-expression and MDR creation and that CAMKIID and CREB1 are attractive targets for restoring doxorubicin efficacy in ABCB1-mediated MDR ovarian cancer.

Tuberculosis (T.B.) is an infectious disease caused primarily by the bacterial pathogen Mycobacterium tuberculosis (Mtb) in humans. The emergence of various drug-resistant Mtb strains threatens to disrupt worldwide attempts to control the infection. The state of disease can be attributed to the host–pathogen protein interaction network. Hence in this study, we have analyzed the protein interaction network at the intra-species level, i.e., within Mtb using the Louvain community detection and at the inter-species level, i.e., between human and Mtb proteins. We observed a higher inter-connectedness in the intra-species protein network as compared to the inter-species network. This reflects a critical role of protein interaction network modulation during host–pathogenesis conditions. After analyzing 269 Mtb proteins and 2287 human proteins using the interlog approach, we observed a total of 640 host–pathogen protein interaction pairing involving 120 humans and 90 Mtb proteins. The H.P.I.s were filtered using functional protein annotations and publicly available experimental results. In the Mtb PPI network, Rv1286, Rv0462, Rv2215, Rv3559c, Rv3504, and Rv3523 are associated as a motif (a governing unit in a network) and identified as key proteins. This interaction network will help researchers better understand host–pathogen protein–protein interactions, as well as shed light on how Mtb interacts with its host.

The leaf of Chinese prickly ash, a unique spice having typical pungent sensation, is a popular food in Southwest China with antipruritic, insecticidal and fungicidal functions, but its bioactive constituents of fungistatic capacity remain unknown. In present investigation, twenty-nine compounds were isolated from leaf of Chinese prickly ash, and their antifungal bioactivity against drug-resistant Candida albicans were evaluated in vitro and in vivo. As a result, three compounds 3, 10, 29 showed antifungal bioactivity by damage of the fungal biofilm, and they might recover sensitive of drug resistant C. albicans to Fluconazole. Then Chinese prickly ash leaf was proved to be a functional food against fungus for the first time in experiment.

The evolution and incidence of multidrug-resistant food-borne pathogens still become a critical public health global issue. To avert this challenge there is great interest in medical applications of silver nanoparticles. Thus, this study aimed to synthesize silver nanoparticles (Rn-AgNPs) using aqueous leaf extract of Nepal Dock (Rumex nepalensis Spreng) and evaluate their antibacterial potential against food-borne pathogens and antioxidant activity. The Rn-AgNPs were characterized by UV-Vis spectrophotometry, Dynamic Light Scattering (DLS), Scanning Electron Microscopy (SEM), and Fourier Transform Infra-Red Spectroscopy (FTIR). The antibacterial activities of the Rn-AgNPs were evaluated using agar well diffusion (zone of inhibition, ZOI) and microdilution (minimum inhibitory concentration, MIC and minimum bactericidal concentration, MBC) methods. The antioxidant property of the Rn-AgNPs was investigated using radical scavenging (DPPH and hydroxyl) assays. The UV-Vis spectra of Rn-AgNPs elucidated the absorption maxima at 425 nm and FTIR detected numerous functional groups of biological compounds that are responsible for capping and stabilizing Rn-AgNPs. DLS analysis displayed monodispersed Rn-AgNPs of 86.7 nm size and highly negative zeta potential (-32.5 mV). Overall results showed that Escherichia coli was the most sensitive organism, whereas Staphylococcus aureus was the least sensitive against Rn-AgNPs. In the antioxidant tests, the AgNPs radical scavenging activity reached 95.44% at 100 μg/ml. This study indicates that Rn-AgNPs exhibit a strong antimicrobial on L. monocytogenes, S. aureus, S. typhimurium, and E. coli and antioxidant and thus might be developed as a new type of antimicrobial agent for the treatment of multidrug-resistant foodborne pathogens and extensible applications in nanomaterial food- and nanocomposite-based antimicrobial packaging and/or as an antioxidant.

Antibiotic resistance in Pseudomonas aeruginosa remains one of the most challenging phenomena of everyday medical science. The universal spread of high-risk clones of multidrug-resistant/extensively drug-resistant (MDR/XDR) clinical P. aeruginosa has become a public health threat. The P. aeruginosa bacteria exhibits remarkable genome plasticity that utilizes highly acquired and intrinsic resistance mechanisms to counter most antibiotic challenges. In addition, the adaptive antibiotic resistance of P. aeruginosa, including biofilm-mediated resistance and the formation of multidrug-tolerant persisted cells, are accountable for recalcitrance and relapse of infections. We highlighted the AMR mechanism considering the most common pathogen P. aeruginosa, its clinical impact, epidemiology, and save our souls (SOS)-mediated resistance. We further discussed the current therapeutic options against MDR/XDR P. aeruginosa infections, and described those treatment options in clinical practice. Finally, other therapeutic strategies, such as bacteriophage-based therapy and antimicrobial peptides, were described with clinical relevance.

Silver nanoparticles (AgNPs) were biosynthesized using the aqueous extract ofMentha pulegium leaves and were characterized by Ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, Scanning electron microscopy, X-ray diffraction, Energy-dispersive X-ray spectroscopy, and X-ray fluorescence analysis. The bactericidal activity was assayed by disc diffusion and broth microdilution methods. The cytotoxicity was examined on normal human peripheral blood mononuclear cells (PBMCs) by MTT assay. SEM and XRD analysis showed the spherical and face-centered cubic nano silver particles with an average size of 29 nm. EDX and XRF spectra exhibited obvious signals in the silver region. The AgNPs displayed antibacterial activity against Staphylococcus aureus followed by Bacillus cereus, Klebsiella pneumoniae and Escherichia coli with minimum inhibitory concentrations (MICs) ranged from 61.5 to 125 µg/mL. No significant cytotoxicity was observed against the PBMCs up to 250 μg/mL of the AgNPs and hence the synthesized AgNPs may be used to treat drug-resistant bacteria in biomedicine. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the effectiveness and tolerability of perampanel (PER) in real-world settings in patients between 1 month and 18 years of age with drug resistant epilepsy (DRE) waiting for epilepsy surgery.Methods: In this multicenter study, patients between 1 month and 18 years of age with DRE treated with PER between January 2020 and June 2021 were selected. The study outcome was effectiveness of PER treatment reported as reduction in seizure frequency and seizure freedom rate. Effectiveness was assessed at 30, 60, 90, 120, 150 and 180 days after initiation of PER. Tolerability profiles were reported as adverse events according to the observations of the patients' family members and physician.Results: Eighty-five patients treated with PER were included in the study. The mean initial dose and mean maximum dose of adjunctive PER was 2 mg/day and 5.8 mg/day, respectively. The mean seizure frequency (rate/week) was 41.3, 25.4, 18.9, 14.3, 11.2, 11.1 and 8.9 seizures at baseline, 30, 60, 90, 120, 150 and 180 days, respectively; the reduction in the mean seizure frequency at all timepoints was significant compared at the baseline (p<0.001). At 180 days, ≥75% seizure reduction was seen in 64.9% (37/57) of the patients and seizure freedom was achieved in 36.8% (21/57). Drowsiness, ataxia, and behavioral changes were the common adverse events observed, and these improved after the dose of PER was reduced. No discontinuation of PER was required due to side effects or intolerance.Conclusion: In real-world settings, PER is well tolerated and effective in seizure control in pediatric and adolescent patients with DRE. [ABSTRACT FROM AUTHOR]

Larrea tridentata antibacterial lignan 4,4′-dihydroxy-3-methoxy-6,7′-cyclolignan (1) was derivatized to obtain eleven new amino ether derivatives (2 A - 12 C). The structural elucidation of compounds was performed by analysis of 1D- and 2D NMR spectral data and HRESIMS. The antibacterial activity of compounds was determined against nine drug-resistant bacteria and two strains of Mycobacterium tuberculosis (sensitive ATCC 27294 H37Rv and drug-resistant G122). Results showed that all derivatives were devoid of activity towards six gram-negative clinical isolates assayed. However, seven derivatives displayed antibacterial activity against three gram-positive drug-resistant bacteria. Further, enhancement of antibacterial activity was only observed for the compounds 2 A and 10 C - 12 C (MIC of 12.5 µg/mL) which were two-fold more active than the starting material 1 against vancomycin-resistant Enterococcus faecium. All derivatives, except compound 9 B , showed antitubercular activity against both M. tuberculosis strains. Interestingly, all the compounds, except for 2 A and 11 A , were more active than the starting material 1 (MIC of 50 µg/mL). Compound 4 C was the only compound as active as the positive control ethambutol against the drug-resistant strain M. tuberculosis G122 (MIC of 6.25 µg/mL). In addition, the derivative 7 C was the most active compound against the sensitive strain M. tuberculosis H37Rv (MIC of 6.25 µg/mL) [Display omitted] • Aminoether analogues of 4,4′-dihydroxy-3-methoxy-6,7′-cyclolignan were synthesized. • The antibacterial activity of the compounds was determined. • 4 C was as active as the control drug ethambutol against a multidrug resistant Mtb strain. [ABSTRACT FROM AUTHOR]

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Epilepsy is a chronic neurological disorder that affects more than 50 million people worldwide. Despite a recent introduction of antiseizure drugs for the treatment of epileptic seizures, one‐third of these patients suffer from drug‐resistant epilepsy (DRE). The therapeutic target hypothesis is a cited theory to explain DRE. According to the target hypothesis, the failure to achieve seizure freedom leads to alteration of the structure and/or function of the antiseizure medication (ASM) target. However, this hypothesis fails to explain why patients with DRE do not respond to antiseizure medications of different targets. This review presents different conditions, such as epigenetic mechanisms and protein‐protein interactions that may result in alterations of diverse drug targets using different mechanisms. These novel conditions represent new targets to control DRE. [ABSTRACT FROM AUTHOR]

Simple Summary: Salmonellosis, an infection in humans and animals caused by Salmonella spp., poses a major concern to public health and food safety worldwide. Antibiotics are mostly prescribed to treat salmonellosis. Unfortunately, indiscriminate use of antibiotics leads to the emergence and transmission of multidrug-resistant Salmonella spp. As antibiotics are becoming increasingly ineffective, infections caused by MDR strains will be difficult to manage. The search for an alternative to antibiotics has led scientists to give renewed attention on phage therapy. Though commercial use of phages for controlling Salmonella in poultry is still in its early stage, the use of lytic phages is considered an environmentally friendly, cost-effective, and sustainable antimicrobial approach. Moreover, it provides advantages over antibiotics in terms of specificity, cost of development, resistance, and genetic amenability. Studies on laboratory and field scale use show promise on the effectiveness of phages against MDR Salmonella spp. However, inadequate data on safety of phage use, phage stability, and lack of regulatory framework remain major obstacles in the commercial application of phages. Our article provides a comprehensive overview on global prevalence and antimicrobial resistance of Salmonella in poultry, the efforts to control Salmonella using phage therapy, and challenges as well as future prospects of phage therapy. Salmonellosis is one of the most common bacterial infections that impacts both human health and poultry production. Although antibiotics are usually recommended for treating Salmonella infections, their misuse results in the evolution and spread of multidrug-resistant (MDR) bacteria. To minimize the health and economic burdens associated with antimicrobial resistance, a novel antibacterial strategy that can obliterate pathogens without any adverse effects on humans and animals is urgently required. Therefore, therapeutic supplementation of phages has gained renewed attention because of their unique ability to lyse specific hosts, cost-effective production, environmentally-friendly properties, and other potential advantages over antibiotics. In addition, the safety and efficacy of phage therapy for controlling poultry-associated Salmonella have already been proven through experimental studies. Phages can be applied at every stage of poultry production, processing, and distribution through different modes of application. Despite having a few limitations, the optimized and regulated use of phage cocktails may prove to be an effective option to combat infections caused by MDR pathogens in the post-antibiotic era. This article mainly focuses on the occurrence of salmonellosis in poultry and its reduction with the aid of bacteriophages. We particularly discuss the prevalence of Salmonella infections in poultry and poultry products; review the trends in antibiotic resistance; and summarize the application, challenges, and prospects of phage therapy in the poultry industry. [ABSTRACT FROM AUTHOR]

Background. The genomic era has led to enormous progress in clinical care and a multi-disciplinary team (MDT) approach is imperative for integration of genomics into epilepsy patient care. Methods. The MDT approach involved patient selection, genomic testing choice, variant discussions and return of results. Genomics analysis included cytogenomic testing and whole exome sequencing (WES). Neurologist surveys were undertaken at baseline and after genomic testing to determine if genomic diagnoses would alter their management, and if there was a change in confidence in genomic testing and neurologist perceptions of the MDT approach. Results. The total diagnostic yield from all genomic testing was 17% (11/66), with four diagnoses from cytogenomic analyses. All chromosomal microarray (CMA) diagnoses were in patients seen by adult neurologists. Diagnostic yield for WES was 11% (7/62). The most common gene with pathogenic variants was DCX, reported in three patients, of which two were mosaic. The genomic diagnosis impacted management in 82% (9/11). There was increased confidence with integrating genomics into clinical care (Pearson chi square = 83, p = 0.004) and qualitative comments were highly supportive of the MDT approach. Conclusions. We demonstrated diagnostic yield from genomic testing, and the impact on management in a cohort with drug-resistant epilepsy. The MDT approach increased confidence in genomic testing and neurologists valued the input from this approach. The utility of CMA was demonstrated in epilepsy patients seen by adult neurologists as was the importance of considering mosaicism for previously undiagnosed patients. [ABSTRACT FROM AUTHOR]

Acute myeloid leukemia (AML) is an aggressive hematological tumor caused by the malignant transformation of myeloid progenitor cells. Although intensive chemotherapy leads to an initial therapeutic response, relapse due to drug resistance remains a significant challenge. In recent years, accumulating evidence has suggested that posttranscriptional methylation modifications are strongly associated with tumorigenesis. However, the mRNA profile of m7G modification in AML and its role in drug-resistant AML are unknown. In this study, we used MeRIP-seq technology to establish the first transcriptome-wide m7G methylome profile for AML and drug-resistant AML cells, and differences in m7G between the two groups were analyzed. In addition, bioinformatics analysis was conducted to explore the function of m7G-specific methylated transcripts. We found significant differences in m7G mRNA modification between AML and drugresistant AML cells. Furthermore, bioinformatics analysis revealed that differential m7Gmodified mRNAs were associated with a wide range of cellular functions. Importantly, down-methylated m7G modification was significantly enriched in ABC transporter-related mRNAs, which are widely recognized to play a key role in multidrug resistance. Our results provide new insights into a novel function of m7G methylation in drug resistance progression of AML. [ABSTRACT FROM AUTHOR]

Drug resistance is one of the problems affecting the world where drug-resistant Escherichia coli and Klebsiella pneumoniae have been shown to be ubiquitous, frequently isolated from foods and commonly associated with surgical site infection in hospitals worldwide. The aims of this work were to analyze the antibacterial activity of root extracts of the plant obtained by serial extraction (using petroleum ether, chloroform, methanol, and water) and by in vitro techniques and preliminary screen phytochemicals present in the extract by qualitative means. Fresh roots of Plumbago indica were collected, oven-dried, and extracted using Soxhlet apparatus; antibacterial activity, minimum inhibitory concentrations (MICs), and minimum bactericidal concentrations (MBCs) of the active extract were evaluated by standard methods against clinically isolated drug-resistant E. coli and K. pneumoniae; preliminary phytochemical screening was taken to detect the presence of alkaloids, saponins, flavonoids, steroids, tannins, reducing sugars, phenolics, protein, and oil and fat; and bioactive compounds were detected by GCMS analysis of the active extracts. Determination of antibacterial activity showed that the test organisms were susceptible to methanol and aqueous extracts only. MIC of methanolic extract was found to be 20 µg/mL on both E. coli and K. pneumoniae, while aqueous extract had MIC of 10 and 20 µg/mL on E. coli and K. pneumoniae, respectively. Preliminary phytochemical screening showed the presence of all the above-mentioned phytochemicals except oil and fat. The significance of this work is to find a lasting solution to the current problem of emerging drug-resistant bacteria (E. coli and K. pneumoniae) through the use of extracts obtained from P. indica which have long history of use as traditional medicine. The methanolic and aqueous extract can be recommended as an alternative and candidates for drug development against drug-resistant E. coli and K. pneumoniae.

The widespread use of broad-spectrum antibiotics, the growing number of immunocompromised individuals, and the emergence of drug-resistant strains have resulted in the increasing incidence and mortality of invasive fungal infections. Azole drugs are the primary treatment for invasive fungal infections, and Iodiconazole is a potent azole drug with strong antifungal activity, but its stability is poor. In order to improve stability, a series of triazole compounds containing ethynyl group were designed and synthesized. Most of the compounds showed strong inhibitory activity against pathogenic fungi, among which compound 20l showed excellent inhibitory activity against pathogenic fungi and drug-resistant fungi. Importantly, and the stability of 20l (T 1/2 = 30.2 min) was obviously improved compared with Iodiconazole (T 1/2 = 4.39 min). In addition, the preferred compound 20l can prevent fungal phase transition and the formation of fungal biofilm, and show satisfactory fungicidal activity. In addition, the compound 20l was almost non-toxic to mammalian HUVEC cell and 293T cell. In vivo pharmacokinetic studies showed that 20l had acceptable pharmacokinetic properties. These results strongly demonstrate that compound 20l was worth further investigation as a potential antifungal inhibitor. [Display omitted] • Compound 20l exhibited excellent antifungal activities against pathogenic fungi. • Compound 20l displayed potent antifungal activities on against FLC-resistant strains. • Compound 20l demonstrated excellent fungicidal activity. • Compound 20l exhibited moderate pharmacokinetic properties. [ABSTRACT FROM AUTHOR]

Invasive fungal infections (IFIs) are emerging as a serious infectious disease worldwide. Due to the lack of effective antifungal drugs and serious drug resistance, the number of people with low immunity is increasing, leading to high morbidity and mortality. Azole drugs targeting CYP51 are widely used in the treatment of invasive fungal infections. By analyzing representative azole antifungal drugs, the characteristics of pharmacophore were summarized. The binding mode of lead compound Iodiconazole was analyzed, and it was found that the narrow hydrophobic cavity was not fully occupied. Therefore, a series of triazole compounds were designed and synthesized by fragment growth strategy. Most of the compounds showed strong inhibitory activity against pathogenic fungi, among which compound A33 showed excellent inhibitory activity against pathogenic fungi and drug-resistant strains. In addition, the preferred compound A33 can prevent fungal phase transition, the formation of fungal biofilm, and show satisfactory fungicidal activity. In addition, the compound A33 was almost non-toxic to mammalian HUVEC cell. These results strongly suggested that compound A33 was worth further investigation as a potential azole inhibitor. [Display omitted] • Based on the Iodiconazole, a series of novel CYP51 inhibitors were designed and synthesized. • Compound A33 exhibited excellent antifungal activities against pathogenic fungi. • Compound A33 displayed potent antifungal activities on against FLC-resistant strain. • Compound A33 inhibited the formation of fungal biofilm. • Compound A33 demonstrated excellent fungicidal activity. [ABSTRACT FROM AUTHOR]

Abstract Epilepsy is a chronic neurological disorder that affects more than 50 million people worldwide. Despite a recent introduction of antiseizure drugs for the treatment of epileptic seizures, one‐third of these patients suffer from drug‐resistant epilepsy (DRE). The therapeutic target hypothesis is a cited theory to explain DRE. According to the target hypothesis, the failure to achieve seizure freedom leads to alteration of the structure and/or function of the antiseizure medication (ASM) target. However, this hypothesis fails to explain why patients with DRE do not respond to antiseizure medications of different targets. This review presents different conditions, such as epigenetic mechanisms and protein‐protein interactions that may result in alterations of diverse drug targets using different mechanisms. These novel conditions represent new targets to control DRE.

ABSTRACT Data regarding the epidemiology of extensively drug-resistant (XDR) carbapenemase-producing Enterobacterales (CPE) in Canada are scarce. Among CPE patients identified by the Canadian Nosocomial Infection Surveillance Program, the following were each significantly associated with XDR status: international travel history; CPE acquisition from a health care exposure abroad; presence of the New Delhi metallo-β-lactamase (NDM) carbapenemase gene; E. coli sequence type (ST) 167, ST405, and ST648; E. cloaceae ST177; C. freundii ST22; and resistance to all antimicrobials except colistin, tigecycline, and ceftazidime-avibactam. IMPORTANCE Extensively drug-resistant (XDR) carbapenemase-producing Enterobacterales (CPE) are a global public health concern. XDR CPE are among the most drug-resistant and difficult-to-treat bacteria, and infected patients are likely to experience adverse outcomes. Because XDR status further reduces effective therapeutic options, it is critical for clinicians to consider resistance and therapeutic options not only in the context of a patient with CPE but also in the context of potential XDR status. Our study reports on patient characteristics associated with the acquisition of an XDR CPE. Our study also reports on the species and carbapenemases associated with XDR status among Enterobacterales identified in Canada. Among a panel of 22 antibiotics, including novel combination drugs, we showed which retained the highest activity against XDR CPE, which may help guide the selection of antibiotic treatments.

The emergence of drug-resistant bacteria, facilitated by metallo-beta-lactamases (MBLs), presents a significant obstacle to the effective use of antibiotics in the management of clinical drug-resistant bacterial infections. AFM-1 is a MBL derived from Alcaligenes faecalis and shares 86% homology with the NDM-1 family. Both AFM-1 and NDM-1 demonstrate the ability to hydrolyze ampicillin and other β-lactam antibiotics, however, their substrate affinities vary, and the specific reason for this variation remains unknown. We present the high-resolution structure of AFM-1. The active center of AFM-1 binds two zinc ions, and the conformation of the key amino acid residues in the active center is in accordance with that of NDM-1. However, the substrate-binding pocket of AFM-1 is considerably smaller than that of NDM-1. Additionally, the mutation of amino acid residues in the Loop3 region, as compared to NDM-1, results in the formation of a dense hydrophobic patch comprised of hydrophobic amino acid residues in this area, which facilitates substrate binding. Our findings lay the foundation for understanding the molecular mechanism of AFM-1 with a high affinity for substrates and provide a novel theoretical foundation for addressing the issue of drug resistance caused by B1 MBLs. • The high-resolution crystal structure of AFM-1 was resolved. • The crystal structure of AFM-1 reveals a smaller substrate binding pocket in comparison to NDM-1. • The decreased K m of AFM-1 for ampicillin can be attributed to the fact that Loop3 forms a more aggregated hydrophobic patch. [ABSTRACT FROM AUTHOR]

• DCS is a technique with high specificity (96.9%) for defining the seizure-onset zone in the presurgical evaluation of drug-resistant epilepsy. • DCS has low sensitivity (23.0%) due to a high percentage of false-negative stimulations obtained during SEEG monitoring. • The resection of the contacts involved in eliciting true positive seizures is associated with a good surgical outcome. Direct cortical stimulation (DCS) is standard for intracranial presurgical evaluation in drug-resistant epilepsy (DRE). Few studies have reported levels of concordance between spontaneous seizure generators and triggered seizures during DCS. The present work reports validity measures of DCS for detecting the seizure onset zone (SOZ) during stereoelectroencephalography (SEEG). We evaluated all patients who underwent SEEG evaluation at our epilepsy center between 2013 and 2019. Data were analyzed using contingency tables. Validity measures of the diagnostic test were computed for all patients evaluated with DCS and for seizure free patients. Fifty-eight consecutive patients were evaluated through DCS. One hundred seventy-three clinical seizures were elicited with DCS. Electroclinical identical to spontaneous seizures were considered true positive (TP) seizures. They showed a high specificity (96.9%) for detecting the SOZ in patients that remained seizure free one year after treatment. Sensitivity was low (23.0%), and a high percentage of false-negative stimulations was documented in the SOZ. The accuracy was 87.9%. DCS is a technique with high specificity but a low sensitivity for the localization of the SOZ. The DCS validity measures need to be known when considered for surgical decisions. The interpretation of DCS-triggered seizures and the differentiation of true-positive vs false-positive seizures should be carefully evaluated. DCS seizure triggering is highly specific for SOZ localization. [ABSTRACT FROM AUTHOR]

Purpose: Drug Resistant -Epilepsy is still a major challenge in pharmacotherapy of epilepsy. Pharmacogenetic pathways are one of the most important elements that can help clinicians determine medication response and provide more efficient drug therapy, especially in cases of drug resistance. Genetic alterations in drug target and transporter proteins, in part, could explain the development of drug-resistant epilepsy. We sought to assess the association of CYP3A5 (rs776746), SCN1A (rs2298771) and ABCG2 (rs2231137) candidate polymorphisms with drug-resistant epilepsy among Iranian children with epilepsy.Methods: In a hospital-based case-control study, 93 participants, including 45 men and 48 women aged 1.5 to 14 years old were recruited. Genotyping of CYP3A5 (rs776746), SCN1A (rs2298771) and ABCG2 (rs2231137) polymorphisms using the high-resolution melting (HRM) method were measured in 46 children with drug-resistant epilepsy and 47 healthy control subjects. The binary logistic regression model was used to estimate the odds ratio (OR) for each polymorphism per effect allele increase.Results: The mean (standard deviation [SD]) age of the drug-resistant patients was 10.7 (9.0) years versus 7.3 (3.6) in the control group. In the case group, most of the patients with epilepsy were diagnosed with generalized seizure (about 87%) and negative epileptic history status (63%). Furthermore, idiopathic epilepsy was dominant in the case group (69%). There was a clinically meaningful increase in the chance of drug-resistant epilepsy in participants with candidate polymorphism in ABCG2 gene (per allele T increase, adjusted odds ratio [OR] 2.41, confidence interval [CI] 0.99 to 5.87, P=0.05). No significant association was found between CYP3A5 (per allele C increase, OR 0.92, CI 0.33 to 2.60, P= 0.88) and SCN1A (per allele *1 increase, OR 0.65, 95% CI 0.34 to 1.23, P= 0.19) with drug-resistant epilepsy.Conclusion: We found evidence for the relationship between the ABCG2 gene polymorphism and a higher chance of drug-resistant epilepsy in children. This finding may have important implications for understanding the role of ABCG2 gene polymorphism in children with drug-resistant epilepsy. [ABSTRACT FROM AUTHOR]

Ovarian cancer (OC) is the most lethal of all gynecologic malignancies with poor survival rates. Although surgical treatment and chemotherapy had advanced to improve survival, platinum-based chemoresistance remains a major hurdle in the clinical treatment of OC. The search for novel active ingredients for the treatment of drug-resistant OC is urgently needed. Here, we demonstrated that icaritin, the main active ingredient derived from the traditional Chinese herb Epimedium genus, significantly suppressed the proliferation, migration, and invasion of both drug-susceptible and cisplatin-resistant OC cells in vitro. Mechanistically, icaritin at 20 μM significantly inhibited the phosphorylation of Akt and mTOR, as well as decreased the expression of vimentin and increased the expression of E-cadherin. Our data indicate that icaritin, a prenylated flavonoid natural product, could serve as a potential inhibitor of cisplatin-resistant OC by inhibiting the Akt/mTOR signaling pathway. [ABSTRACT FROM AUTHOR]

Up to now, it has not been clear whether occult hepatitis B virus (HBV) infection (OBI) can be treated with antiviral therapy whether OBI can develop drug resistance gene mutation or not. We report a middle-aged female patient with OBI who showed HBV reactivation (HBVr) during more than 3 years of intermittent entecavir (ETV) antiviral therapy: seropositive HBV surface antigen (HBsAg), increased e antigen (HBeAg), and repeatedly elevated serum HBV DNA. Genotype analysis showed that the patient was infected with HBV type B. Genetic sequencing of HBV showed the mutants of S143T, D144G, and G145R in the S gene region, and the mutant of site 1896 in the pre-Core region coexisted with the wild type (G1896A/G). No mutation was found in other HBV gene segments. Drug resistance gene analysis found RtL229W mutant, resistant to lamivudine but sensitive to ETV and other nucleoside analogs. This case of OBI provides us with the following clinical experiences: Firstly, it is necessary to detect HBV genotype, mutation, and drug-resistant genes at the initial diagnosis, which can be helpful for reasonable treatment. Secondly, identifying the risk factors and mechanisms associated with HBVr could help quantify the risk of HBVr and manage the clinical consequences. Thirdly, the OBI patients with hepatitis B e antigen-positive, HBV DNA > 1 × 103 IU/ml should be recommended regular and continuous antiviral therapy as soon as possible to prevent the occurrence of hepatocirrhosis and hepatocellular carcinoma (HCC). [ABSTRACT FROM AUTHOR]

The primary aim of this study was to evaluate the efficacy of phage against mastitis induced by drug-resistant S. aureus in a mouse model. In this study, five S. aureus phages—4086-1, 4086-2, 4086-3, 4086-4, and 4086-6—were isolated from milk samples secreted by mastitis cows. Transmission electron microscopy showed that all the five phages had icosahedral heads and short non-contractile tails, which are typical characteristics of the family Podoviridae. All these phages were species-specific against S. aureus. The one-step growth curve showed a short latency period (10–20 min) and high burst size (up to 400 PFU/infected cell). To evaluate the effectiveness of the phage 4086-1 in the treatment against mastitis, a mouse model of mastitis was challenged with drug-resistant S. aureus. The results showed the proliferation of S. aureus in the mammary glands was significantly inhibited after treating by phage 4086-1. The concentrations of TNF-α and IL-6 decreased significantly, which demonstrated the phages could effectively alleviate the inflammatory responses. Furthermore, the histopathological analysis showed that inflammatory infiltration in the mammary glands was significantly reduced. These results demonstrate that phage may be a promising alternative therapy against mastitis caused by drug-resistant S. aureus. [ABSTRACT FROM AUTHOR]

Helicobacter pylori (H. pylori) infection is prevalent all over the world, which may even cause gastric cancer. Unfortunately, current antibiotic therapy cannot completely eradicate drug-resistant H. pylori. HF-18 (GFFKKAWRKVKKAFRRVL-NH2), a novel cationic antimicrobial peptide, showed broad antimicrobial activities, low cytotoxicity toward mammalian cells and low drug tolerance during our previous research. In this paper, to investigate the antibacterial activities and possible mechanisms underlying HF-18 against drug-resistant H. pylori, cytotoxin associated protein (CagA)-carrying recombinant E. coli BL21 was constructed. Subsequently, Zeta potential detection, N-Phenyl-1-naphthylamine uptake, propidium iodide fluorescence uptake, transmission electron microscope, qRT-PCR and Western blot assays were performed. Consequently, we found that HF-18 had potent antibacterial effects on clarithromycin- and amoxicillin- resistant H. pylori. In terms of its mode of action, HF-18 could neutralize negative charges on the surface of H. pylori, penetrate into and disrupt the integrity of bacterial membranes, resulting in the death of drug-resistant H. pylori to reduce cells infection. In addition, HF-18 inhibited the transcription of adhesion gene alpA/alpB and the expression of virulence protein CagA under sub-inhibitory concentration, and ultimately decreased bacterial virulence. Therefore, HF-18 lays a foundation for the development and utilization of new antibiotics. [ABSTRACT FROM AUTHOR]

Abstract Background Multidrug-resistant tuberculosis (MDR-TB) in HIV infected individuals is a serious threat to global efforts to combat tuberculosis. Inconsistent findings on the association between HIV infection and MDR-TB were present in many studies. We aimed to review existing data on the relationship between HIV infection and MDR-TB systematically to assess the contribution of HIV on MDR-TB worldwide. We also investigated the patterns of MDR-TB by age, country-wise income, study designs, and global regions. Methods We utilized PubMed, Google Scholar, and ScienceDirect databases to select eligible studies for meta-analysis that were published between January 12,010, and July 30, 2020. The random-effects model was used to obtain the pooled odds ratio of the crude association between HIV and MDR-TB with a 95% confidence interval. We investigated the potential publication-bias by checking funnel plot asymmetry and using the Egger’s test. Moreover, we assessed the heterogeneity using the I 2 statistic. Sensitivity analysis was performed based on sample size and adjustment factors. The protocol was registered with PROSPERO-CRD42019132752. Results We identified 1603 studies through a database search, and after subsequent eliminations we selected 54 studies including 430,534 TB patients. The pooled odds of MDR-TB was 1.42 times higher in HIV-positive patients than HIV-negative patients (OR=1.42,CI=1.17–1.71, I 2 =75.8%). Subgroup analysis revealed that the estimated pooled odds for South-East Asian countries was 1.86, which is the highest in WHO regions (OR=1.86,CI=1.30–2.67, I 2 =0.00%), followed by Europe and Africa. The effect estimate was found to be higher for primary MDR-TB (OR=2.76,CI=1.70–4.46, I 2 =0.00%). There was also a trend towards increased odds of MDR-TB for HIV patients older than 40 years (OR=1.56,CI=1.17–2.06). The association was found to be significant in high-burden TB countries (OR=1.75, CI=1.39–2.19) and in high-income countries (OR=1.55, CI=1.06–2.27). Conclusion Such findings indicate that HIV infection raises the risk of MDR-TB, and after contrasting it with the results of the earlier pooled study, it appeared to be an upward risk trend. Moreover, we found that the risk is the highest in the South-East Asian region. A balanced allocation of resources is needed to halt both primary and secondary MDR-TB, particularly in HIV infected people with 40 years of age and older.

Context Yang-Yin-Jie-Du Decoction (YYJDD) was used to improve gefitinib efficacy in our clinical practice, but its mechanism remains unclear. Objective This study explored if YYJDD could reverse gefitinib resistance. Materials and methods H1975 cells were exposed to control, 10 μM gefitinib, 3.2 mg/mL YYJDD or combination treatment. Cell viability was detected by MTT during 0–96 h. Apoptosis and the PI3K/Akt proteins were tested by flow cytometry and western-blot at 24 h. LY294002 was applied to further determine the role of the PI3K/Akt. 23 BALB/c nude xenograft mice received normal saline (n = 5), 80 mg/kg gefitinib (n = 6), 2.35 g/kg lyophilised powder of YYJDD (n = 6) or combination treatment (n = 6) by gavage for 4 weeks and submitted to TUNEL, immunohistochemistry, and western-blot. Results In vitro, gefitinib (IC50: 20.68 ± 2.06 μM) and YYJDD (IC50: 6.6 ± 0.21 mg/mL) acted in a moderate synergistic way. Combination treatment inhibited cell viability from 100% to 25.66%. Compared to gefitinb (33.23 ± 3.99%), cell apoptosis was increased with combination treatment (54.11 ± 7.32%), accompanied by down-regulation of the PI3K/Akt. LY294002 further inhibited cell viability, increased apoptosis, and down-regulated p-Akt/Akt. In vivo, the tumour sizes in the combination group (1165.13 ± 157.79 mm3) were smaller than gefitinib alone (1630.66 ± 208.30 mm3). The positive rate of TUNEL staining was increased by combination treatment (22.33 ± 2.75%) versus gefitinib (7.37 ± 0.87%), while the PI3K/Akt was down-regulated. Discussion and conclusion YYJDD has potential to overcome gefitinib resistance. Future investigations should be focussed on its specific targets.

Bladder cancer is the most common malignant tumor of the urinary system with high morbidity and no clear pathogenesis. The Hippo signaling pathway is an evolutionarily conserved pathway that regulates organ size and maintains tissue homeostasis. Yes‑associated protein 1 (YAP1) is a key effector of this pathway and regulates downstream target genes by binding to transcriptional co‑activators with PDZ binding sequences (TAZ). Several studies have demonstrated that YAP1 is overexpressed in bladder cancer and is involved in adverse outcomes such as bladder cancer occurrence, progression, resistance to cisplatin and the recurrence of tumours. The present review summarized the involvement of YAP1 in bladder cancer disease onset and progression, and the mechanism of YAP1 involvement in bladder cancer treatment. In addition, this study further explored the potential of YAP1 in the diagnosis and treatment of bladder cancer. This study aimed to explore the potential mechanism of YAP1 in the treatment of bladder cancer.

Vibrio cholerae is the causative organism of the cholera epidemic, and it remains a serious global health problem, particularly the multidrug-resistant strain, despite the development of several generic drugs and vaccines over time. Natural products have long been exploited for the treatment of various diseases, and this study aimed to evaluate the in vitro antibacterial activity of coffee beans and coffee by-products against V. cholerae antimicrobial resistant strains. A total of 9 aqueous extracts were investigated, including light coffee (LC), medium coffee (MC), dark coffee (DC), dried green coffee (DGC), dried red coffee (DRC), fresh red coffee (FRC), Arabica leaf (AL), Robusta leaf (RL), and coffee pulp (CP). The influential coffee phytochemicals, i.e., chlorogenic acid (CGA), caffeic acid (CA), and caffeine, were determined using HPLC. The antibacterial properties were tested by agar well-diffusion techniques, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were further determined against 20 V. cholerae isolates. The results revealed that all tested strains were sensitive to coffee extracts, with MIC and MBC values in the range of 3.125–25.0 mg/mL and 12.5–50.0 mg/mL, respectively. With a MIC of 6.25 mg/mL, DGC, DRC, and CP appeared to be the most effective compounds against 65, 60, and 55% of clinical strains, respectively. The checkerboard assay revealed that the combination of coffee extract and tetracycline was greater than either treatment alone, with the fractional inhibitory concentration index (FICI) ranging from 0.005 to 0.258. It is important to note that CP had the lowest FICI (0.005) when combined with tetracycline at 60 ng/mL, which is the most effective dose against V. cholerae six-drug resistance strains (azithromycin, colistin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim), with a MIC of 47.5 μg/mL (MIC alone = 12.5 mg/mL). Time killing kinetics analysis suggested that CA might be the most effective treatment for drug-resistant V. cholerae as it reduced bacterial growth by 3 log10 CFU/mL at a concentration of 8 mg/mL within 1 h, via disrupting membrane permeability, as confirmed by scanning electron microscopy (SEM). This is the first report showing that coffee beans and coffee by-product extracts are an alternative for multidrug-resistant V. cholerae treatment.

Ovarian cancer (OC) is the most lethal of all gynecologic malignancies with poor survival rates. Although surgical treatment and chemotherapy had advanced to improve survival, platinum-based chemoresistance remains a major hurdle in the clinical treatment of OC. The search for novel active ingredients for the treatment of drug-resistant OC is urgently needed. Here, we demonstrated that icaritin, the main active ingredient derived from the traditional Chinese herb Epimedium genus, significantly suppressed the proliferation, migration, and invasion of both drug-susceptible and cisplatin-resistant OC cells in vitro. Mechanistically, icaritin at 20 μM significantly inhibited the phosphorylation of Akt and mTOR, as well as decreased the expression of vimentin and increased the expression of E-cadherin. Our data indicate that icaritin, a prenylated flavonoid natural product, could serve as a potential inhibitor of cisplatin-resistant OC by inhibiting the Akt/mTOR signaling pathway.

Up to now, it has not been clear whether occult hepatitis B virus (HBV) infection (OBI) can be treated with antiviral therapy whether OBI can develop drug resistance gene mutation or not. We report a middle-aged female patient with OBI who showed HBV reactivation (HBVr) during more than 3 years of intermittent entecavir (ETV) antiviral therapy: seropositive HBV surface antigen (HBsAg), increased e antigen (HBeAg), and repeatedly elevated serum HBV DNA. Genotype analysis showed that the patient was infected with HBV type B. Genetic sequencing of HBV showed the mutants of S143T, D144G, and G145R in the S gene region, and the mutant of site 1896 in the pre-Core region coexisted with the wild type (G1896A/G). No mutation was found in other HBV gene segments. Drug resistance gene analysis found RtL229W mutant, resistant to lamivudine but sensitive to ETV and other nucleoside analogs. This case of OBI provides us with the following clinical experiences: Firstly, it is necessary to detect HBV genotype, mutation, and drug-resistant genes at the initial diagnosis, which can be helpful for reasonable treatment. Secondly, identifying the risk factors and mechanisms associated with HBVr could help quantify the risk of HBVr and manage the clinical consequences. Thirdly, the OBI patients with hepatitis B e antigen-positive, HBV DNA > 1 × 103 IU/ml should be recommended regular and continuous antiviral therapy as soon as possible to prevent the occurrence of hepatocirrhosis and hepatocellular carcinoma (HCC).

Juvenile myoclonic epilepsy (JME) is one of the most common epileptic syndromes; it is estimated to affect 1 in 1,000 people worldwide. Most people with JME respond well to medication, but up to 30% of them are drug-resistant. To date, there are no biomarkers for drug resistance in JME, and the poor response to medications is identified in retrospect. People with JME have frontal dysfunction manifested as impaired attention and difficulties in inhibiting habitual responses and these dysfunctions are more pronounced in drug-resistant individuals. Frontal networks play an important role in walking and therefore, gait can be used to overload the neural system and expose subtle changes between people with drug-responsive and drug-resistant JME. Electroencephalogram (EEG) is a promising tool to explore neural changes during real-time functions that combine a cognitive task while walking (dual tasking, DT). This exploratory study aimed to examine the alteration in electrical brain activity during DT in people with drug-responsive and drug-resistant JME. A total of 32 subjects (14 males and 18 females) participated: 11 drug-responsive (ages: 31.50 ± 1.50) and 8 drug-resistant (27.27 ± 2.30) people with JME, and 13 healthy controls (29.46 ± 0.69). The participants underwent EEG examination during the performance of the visual Go/NoGo (vGNG) task while sitting and while walking on a treadmill. We measured latencies and amplitudes of N2 and P3 event-related potentials, and the cognitive performance was assessed by accuracy rate and response time of Go/NoGo events. The results demonstrated that healthy controls had earlier N2 and P3 latencies than both JME groups (N2: p = 0.034 and P3: p = 0.011), however, a limited ability to adjust the N2 amplitude during walking was noticeable in the drug-resistant compared to drug-responsive. The two JME groups had lower success rates (drug-responsive p < 0.001, drug-resistant p = 0.004) than healthy controls, but the drug-resistant showed longer reaction times compared to both healthy controls (p = 0.033) and drug-responsive (p = 0.013). This study provides the first evidence that people with drug-resistant JME have changes in brain activity during highly demanding tasks that combine cognitive and motor functions compared to people with drug-responsive JME. Further research is needed to determine whether these alterations can be used as biomarkers to drug response in JME. [ABSTRACT FROM AUTHOR]